WO1995024490A1 - Reactif de liaison pour proteine de surface cellulaire et cellule effectrice - Google Patents

Reactif de liaison pour proteine de surface cellulaire et cellule effectrice Download PDF

Info

Publication number
WO1995024490A1
WO1995024490A1 PCT/EP1995/000843 EP9500843W WO9524490A1 WO 1995024490 A1 WO1995024490 A1 WO 1995024490A1 EP 9500843 W EP9500843 W EP 9500843W WO 9524490 A1 WO9524490 A1 WO 9524490A1
Authority
WO
WIPO (PCT)
Prior art keywords
binding
binding reagent
reagent according
expression
antibody
Prior art date
Application number
PCT/EP1995/000843
Other languages
German (de)
English (en)
Inventor
Volker Schirrmacher
Khashayarsha Khazaie
Claudia Haas
Gerd Moldenhauer
Melvyn Little
Stefan Dübel
Frank Breitling
Sergey Kipriyanov
Stefanie Gotter
Hans-Jürgen RODE
Original Assignee
Deutsches Krebsforschungszentrum Stiftung des öffentlichen Rechts
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Deutsches Krebsforschungszentrum Stiftung des öffentlichen Rechts filed Critical Deutsches Krebsforschungszentrum Stiftung des öffentlichen Rechts
Priority to JP7523228A priority Critical patent/JPH09510089A/ja
Priority to EP95911308A priority patent/EP0749487A1/fr
Priority to US08/702,712 priority patent/US5911987A/en
Publication of WO1995024490A1 publication Critical patent/WO1995024490A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0005—Vertebrate antigens
    • A61K39/0011—Cancer antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/08—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses
    • C07K16/10—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses from RNA viruses
    • C07K16/1027—Paramyxoviridae, e.g. respiratory syncytial virus
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
    • C07K16/2818—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against CD28 or CD152
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/60—Medicinal preparations containing antigens or antibodies characteristics by the carrier linked to the antigen
    • A61K2039/6031—Proteins
    • A61K2039/6056—Antibodies

Definitions

  • the invention relates to a binding reagent, a process for its preparation and a vaccine containing the binding reagent.
  • NDV Disease Virus
  • the invention is therefore based on the object of providing a means by which the immunogenicity of cells can be increased.
  • this is achieved by providing a binding reagent which is characterized in that it has a first binding component for a cell surface protein and a second binding component for a molecule of an effector cell which has a costimulatory effect.
  • first binding component encompasses any compound which is attached to a cell surface protein on the one hand and to a second binding component on the other can bind component.
  • the binding can be direct or indirect.
  • the compound is an antibody or a portion thereof which has a binding domain.
  • Such a part is favorably a Fab ', (Fab') 2 , F v or (F v ) 2 fragment.
  • the F v fragment of an anti-hemaglutinin neuraminidase antibody has proven to be particularly favorable.
  • cell surface protein encompasses any molecule that may be present on a cell surface. Such can e.g. be a peptide or protein derived from the cell itself, from a virus in the cell or from the expression of a DNA introduced into the cell.
  • the "cell surface protein” is preferably a virus protein, such as the hemaglutinin neuraminidase molecule from NDV, the hemaglutinin molecule from influenza virus or the envelope protein from HTLV-I or HIV, a growth factor receptor, an oncogene product , such as v-Erb B2, an adhesion molecule, an antibody, such as an antibody directed against hapten 2-phenyloxazol-5-one, or streptavidin (avidin) or a part thereof. It has proven to be particularly favorable if the cell surface protein is a virus protein from NDV, with the hemaglutinin neuraminidase molecule from NDV in particular being worth mentioning.
  • a virus protein such as the hemaglutinin neuraminidase molecule from NDV, the hemaglutinin molecule from influenza virus or the envelope protein from HTLV-I or HIV, a growth factor receptor, an oncogene product , such as v-Erb B2, an
  • second binding component encompasses any compound which can bind on the one hand to a first binding component and on the other hand to a costimulatory molecule of an effector cell.
  • the binding can be direct or indirect.
  • the compound is an antibody or a portion thereof which has a binding domain.
  • Such a part is favorably a Fab ', (Fab') 2 , F v or (F v ) 2 fragment.
  • the above compound is a B7 protein or a part thereof (cf. Nature 366 (1 993), 76; Science 262 (1 993), 909 "). It has proven to be particularly advantageous if such Part does not include the membrane-bound domain, and it is also advantageous if the compound is a lymphokine, interferon or interleukin.
  • effector cell encompasses any cell involved in an immune response. It is preferably a T cell.
  • costimulatory molecule includes any molecule of an effector cell that can be caused by binding or otherwise to stimulate the effector cell. Such a molecule can e.g. be a receptor.
  • a receptor e.g. be a receptor.
  • the receptors CD2-, CD3-, CD19-, CD20-, CD22-, CD26-, CD28- and CTLA-4 as well as HSA (Heat Stable Antigen) are particularly advantageous for the invention .
  • the receptor CD28 should be particularly emphasized.
  • the two binding components can be directly or indirectly connected to each other. In the latter case this can be done via a complex, e.g. streptavidin (avidin) and biotin or S-protein and S-peptide of pancreatic ribonuclease A can be brought about.
  • a complex e.g. streptavidin (avidin) and biotin or S-protein and S-peptide of pancreatic ribonuclease A can be brought about.
  • one of the two binding components streptavidin (avidin) or S-protein or a part thereof and the other binding component biotin or S-peptide or a part thereof.
  • Methods are known to the person skilled in the art to couple the constituents of the individual complexes to the binding components and to implement these with one another to form the complexes. Binding reagents in which the binding components are indirectly connected to one another are preferred.
  • binding reagents are preferred in which one or both binding components contain groups which form the formation of intermolecular disulfide bridges, i.e. of disulfide bridges between binding components of different binding reagents.
  • binding reagents that bind different, costimulatory molecules, coupled to a single cell surface protein. In many cases, this proves to be particularly beneficial for increasing the immunogenicity of cells.
  • binding reagents can be prepared by conventional methods.
  • a method is favorable which comprises the following steps: (a) insertion of a DNA encoding a first binding component into an expression vector, expression of the DNA, isolation of the expression product and its purification,
  • Expression vector expression of the DNA, isolation of the expression product and its purification
  • a first binding component and a second binding component are produced by conventional DNA recombination techniques.
  • Systems which can be used to express the individual binding components are known to the person skilled in the art. He knows vectors
  • Expression control elements and cells that can be used for this. It has proven to be advantageous to provide both binding components as F v fragments of antibodies.
  • the expression plasmids pOPE51 - ⁇ HN and pOPE51 - ⁇ CD28 are constructed (see Example 1 below).
  • the expression plasmids pOPE51 - ⁇ HN and pOPE51 - ⁇ CD28 belong to the present invention.
  • the binding component of (a) is coupled with that of (b).
  • Methods which can be used for this are known to the person skilled in the art. He knows those for direct as well as indirect coupling, eg via a streptavidin (Avidin) / biotin or S-protein / S-peptide complex.
  • vidin streptavidin
  • S-protein / S-peptide complex As an example, reference is made to the coupling of the F v - ⁇ HN fragment with the F v - ⁇ CD28 fragment in Example 3 below.
  • Another method has proven to be favorable for producing a binding reagent above, for example one whose binding components F v fragments are of different specificities (specificity A; specificity B).
  • the F v fragments are produced by combining their respective V H and V L domains, some of which are individually expressed.
  • groups which are suitable for the formation of disulfide bridges, for example cysteines are introduced into the domains in such a way that predominantly V H and V L domains of a specificity combine with one another.
  • two mutually compatible expression plasmids are used with which at the same time in a host, e.g. E.coli, the following constructs can be expressed:
  • a vaccine with inactivated cells is also provided, which is characterized in that one or more binding reagents are bound to a cell surface protein.
  • a vaccine preferably has Tumor cells. These can originate from tumors removed by surgery or from an established cell line.
  • the (tumor) cells can be virus-modified. Lysates of (tumor) cells obtained by virus can also be present. NDV from virus is advantageously used. It has proven to be advantageous if the binding reagent (s) are directed against a virus protein from NDV, in particular the hemaglutinin-neuraminidase molecule. As an example, reference is made to the production of the tumor vaccine from Example 4 below.
  • the (tumor) cells can also be expressed by expressing an inserted, e.g. DNA modified for streptavidin (avidin) or an antibody, such as an antibody directed against the hapten 2-phenyloxazol-5-one. It has proven to be advantageous if the binding reagent (s) for streptavidin (avidin) are biotinylated, while for the antibody they contain the hapten 2-phenyloxazol-5-one.
  • binding reagents according to the invention which are to be attached to (tumor) cells, it is possible to transmit signals to molecules having a costimulatory effect, e.g. Receptors to transmit from effector cells. Effector cells thus not only receive the signal mediated by an antigen of (tumor) cells, but are also costimulated.
  • the binding reagents according to the invention are therefore particularly suitable for increasing the immunogenicity of cells, in particular tumor cells. They are a great improvement for active immunization.
  • Fig. 1 shows a schematic representation of the expression plasmid pOPE51 - ⁇ HN.
  • Ap R Ampicillin resistance coding gene
  • bp base pairs
  • c-myc sequence which codes for an epitope which is recognized by mAb 9E10
  • Cys nucleotides encoding a single cysteine residue
  • (His) 5 sequence for five C-terminals Encoded histidine residues
  • IG "intergenic" region of phage f1, leader: signal peptide sequence of the bacterial pectate lyase (pelB leader); left: sequence coding for (Gly 4 Ser) 3 that connects V H and V L
  • ori starting point of DNA replication for ColE1
  • P / O lac operon promoter / operator
  • V H and V L variable region of the heavy or light chain of the anti-HN antibody.
  • FIG. 2 shows a schematic representation of the expression plasmid pOPE51- ⁇ CD28.
  • Ap R for the gene coding for ampicillin resistance; bp: base pairs; c-myc: sequence which codes for an epitope which is recognized by mAb 9E10; Cys: nucleotides coding for a single cysteine residue; (His) 5 : sequence coding for five C-terminal histidine residues; IG: "intergenic" region of phage f1; leader: signal peptide sequence of bacterial pectate lyase (pelB leader); left: sequence coding for (Gly 4 Ser) 3 that connects V H and V L ; ori: position of DNA replication for CoLEI; P / O, lac operon promoter / operator; V and V L : variable region of the heavy and light chain of the anti- ⁇ CD28 antibody.
  • FIG. 3 shows a schematic representation of a binding reagent according to the invention.
  • anti HN anti-HN antibody
  • anti CD28 anti-CD28 antibody
  • S S: disulfide bridge.
  • Example 1 Construction of the expression plasmids pOPE51 - ⁇ HN and pOPE51- ⁇ CD28
  • the vector pOPE40 was used as the starting material (cf. Dübel et al., Gene 128 (1 993), 97). This contains a DNA which codes for the F v fragment (V H + V L ) of an anti-lysozyme antibody. The DNA for V H is linked to that for V L via a linker. The DNA above fragment is followed in the 3 'direction by a DNA which codes for an epitope of the monoclonal antibody 9E10 on the myc gene product.
  • the DNA sequence TGC ATA CAT CAC CAT CAT CAT was inserted at the 3 'end of the above myc DNA from pOPE40.
  • the codon for the amino acid Asn at the 3 'end of the myc DNA was replaced with this DNA coding for the amino acid sequence Cys His His His His.
  • the restriction sites BssHII and EcoRI were removed in the linker between V H and V L.
  • three restriction sites were inserted into the vector portion, namely Ncol (nt. 76-81), Mini (nt. 579-584) and Notl (nt. 910-917).
  • the expression plasmid pOPE51 - ⁇ Lys was obtained.
  • pOPE51 - ⁇ CD28 was carried out starting from pOPE51 - ⁇ Lys, as described under (A) for pOPE51 - ⁇ HN.
  • the DNA coding for the F v fragment (V H + V L ) of an anti-CD28 antibody was extracted from the cDNA one
  • Hybridoma cell line obtained using conventional PCR technology (cf. van Lier, R. et al. In Leucocyte Typing IV, Oxford University Press (1 989), 353).
  • the DNA of this fragment is referred to below as F v - ⁇ CD28-DNA.
  • the expression plasmid pOPE51 - ⁇ CD28 obtained is shown in FIG. 2.
  • E.coli JM 109 cells were transformed in the usual way with the expression plasmid pOPE51 - ⁇ HN and in 2 I LB medium at 30 ° C. up to an OD 600 of
  • Isopropyl- ⁇ -thiogalactoside IPTG was added up to a final concentration of 20 ⁇ m.
  • the cells were incubated at room temperature for 3 h and then collected by centrifugation (4 500 g, 4 ° C., 1 5 min).
  • the cells were suspended in 1/50 of the original volume in 0.1 M sodium acetate, pH 5.5, 10 mM EDTA, 1 mM phenylmethylsulfonyl fluoride (PMSF) at 0 °.
  • IPTG Isopropyl- ⁇ -thiogalactoside
  • Lysozyme was added to a final concentration of 1 mg / ml. After incubation on ice for 30 minutes with gentle shaking, soluble cell plasma proteins were removed by centrifugation (30,000 g, 4 ° C., 30 min). The cell pellets were resuspended in 1/40 of the original volume in 30 mM sodium phosphate, 0.3 M NaCl, 1 mM PMSF, pH 7.0 and lysed by sonication.
  • Imidazole was added to a final concentration of 30 mM.
  • the protein solution was placed on a chelating Sepharose Fast Flow column loaded with NiCl 2 and equilibrated with 6 M urea, 25 M Tris-HCl and 50 mM imidazole, pH 7.0.
  • the column was washed with five times the column volume of 6 M urea, 25 mM Tris-HCl, 50 mM imidazole, pH 7.0.
  • Bound F v - ⁇ HN fragments were eluted with 1.5 times the column volume of 6 M urea, 25 mM Tris-HCl, 250 mM imidazole, pH 7.0.
  • An IMAC was then carried out at room temperature.
  • the eluted protein was dialyzed at 4 ° C against 0.4 M L-arginine-HCl, 0.1 M Tris-HCl, 5 mM EDTA, pH 7.0, then using permeable Collodione-
  • Fat and connective tissue were removed from freshly operated tumor tissue in the usual way.
  • the tumor tissue was cut into small pieces and incubated with 40 ml of an enzyme cocktail (collagenase 0.32 mg / ml, DN-ase 0.535 mg / ml and hyaluronidase 0.535 mg / ml in HBSS) for 1 h at 37 ° C. with stirring.
  • the cell suspension obtained was poured off over a customary nylon mesh.
  • tissue residues were incubated a second time with the above enzyme cocktail (40 ml) and filtered. All suspensions were combined, made up to 50 ml with HBSS and centrifuged for 15 minutes at 1200 rpm. The supernatant was discarded and the pellet was resuspended in 10 ml of the above DN-ase solution and incubated at 37 ° C. for 10 min. The suspension was made up to 50 ml
  • HBSS HBSS filled up and centrifuged at 1300 rpm for 10 min. The pellet was resuspended in 10 ml HBSS and the cells were counted in a Neubauer cell chamber with Trypan blue.
  • Non-tumor cells (lymphocytes and monocytes) of more than 50% of the total cells were a further separation step for removing the lymphocytes and Monocytes performed.
  • 10 Oyc l of Dynabeads anti-CD2 (Pan-T), anti-CD 19 (Pan-B) and anti-CD14 (Pan monocytes) were placed in tubes and washed three times with 7 ml of chilled HBSS / HSA. Cells resuspended in 2 ml of cold HBSS / HSA were added to these tubes and the tubes were incubated on ice for 30 min.
  • Magnetic beads were removed with a magnet and the cell suspension was suctioned off, filled into tubes with 50 ml of HBSS and centrifuged for 10 min at 1300 rpm. The supernatant was discarded, the pellet resuspended in 50 ml HBSS and then centrifuged for 10 min at 1300 rpm.
  • the cell pellet obtained was resuspended in HBSS, 1 ⁇ 10 7 cells being taken up in 0.5 ml HBSS. Approx. 0.5 ml of the cell suspension obtained was placed in prepared freezing tubes. About 0.5 ml of double freezing medium on ice was added to these before the freezing tubes were stored at -70 ° C. overnight and then kept in liquid nitrogen.
  • the above cell suspension was thawed at 37 ° C., transferred to a tube and made up to 14 ml with PBS. After centrifugation at 1300 rpm for 10 minutes, the supernatant was discarded. The pellet was resuspended in 100 l of PBS, transferred to freezing tubes and mixed with 30 ml of NDS suspension (concentration 1000 HAU / ml). This was followed by a 30 minute incubation at 37 ° C. was shaken. After the incubation, washing was carried out carefully before an aliquot of tumor cells (5 ⁇ 10 6 vital tumor cells) was incubated at 37 ° C.
  • Example 3 For 30 min with about 5 // g protein of the binding reagent according to the invention from Example 3. The Zeil suspension was then irradiated with 200 Gy. The Zeil suspension was stored on ice until injection, then drawn into a syringe and injected intradermally with a 0.9x40 ml cannula.

Abstract

L'invention concerne un réactif de liaison qui se caractérise en ce qu'il comporte un premier constituant de liaison pour une protéine de surface cellulaire et un second constituant de liaison pour la molécule à effet costimulateur d'une cellule effectrice. L'invention concerne en outre un procédé de préparation dudit réactif de liaison, ainsi qu'un vaccin contenant ledit réactif de liaison.
PCT/EP1995/000843 1994-03-07 1995-03-07 Reactif de liaison pour proteine de surface cellulaire et cellule effectrice WO1995024490A1 (fr)

Priority Applications (3)

Application Number Priority Date Filing Date Title
JP7523228A JPH09510089A (ja) 1994-03-07 1995-03-07 細胞表層蛋白質及びエフェクター細胞に対する結合試薬
EP95911308A EP0749487A1 (fr) 1994-03-07 1995-03-07 Reactif de liaison pour proteine de surface cellulaire et cellule effectrice
US08/702,712 US5911987A (en) 1994-03-07 1995-03-07 Cell surface protein and effector cell bonding reagent

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DEP4407538.3 1994-03-07
DE4407538A DE4407538C1 (de) 1994-03-07 1994-03-07 Bindungsreagens für Zell-Oberflächenprotein und Effektorzelle

Publications (1)

Publication Number Publication Date
WO1995024490A1 true WO1995024490A1 (fr) 1995-09-14

Family

ID=6512067

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP1995/000843 WO1995024490A1 (fr) 1994-03-07 1995-03-07 Reactif de liaison pour proteine de surface cellulaire et cellule effectrice

Country Status (5)

Country Link
US (1) US5911987A (fr)
EP (1) EP0749487A1 (fr)
JP (1) JPH09510089A (fr)
DE (1) DE4407538C1 (fr)
WO (1) WO1995024490A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1275724A1 (fr) * 2001-07-10 2003-01-15 Volker Prof. Dr. Schirrmacher Réactifs de liaison entre des protéines de surface cellulaire et des cellules effectrices

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB9607711D0 (en) * 1996-04-13 1996-06-19 Univ Sheffield T-cell dependent vaccine
DE19725586C2 (de) * 1997-06-17 1999-06-24 Gsf Forschungszentrum Umwelt Verfahren zur Herstellung von Zellpräparaten zur Immunisierung mittels heterologer intakter bispezifischer und/oder trispezifischer Antikörper
EP1092439A1 (fr) * 1999-10-13 2001-04-18 Thorsten Dr. Ahlert Activation de cellules T antigène spécifiques par des cellules dendritiques traitées par un virus/antigène
CA2858359C (fr) * 2006-11-01 2018-04-03 Ventana Medical Systems, Inc. Haptenes, conjugues de haptene, compositions de haptene, procede de fabrication et utilisation

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0610046A2 (fr) * 1993-02-01 1994-08-10 Bristol-Myers Squibb Company Vecteurs d'expression codant pour des proteins de fusion bispécifiques et méthodes pour la production de ces protéins dans des cellules de mammifèrres
WO1994021798A1 (fr) * 1993-03-16 1994-09-29 British Technology Group Limited Stimulation d'une reponse immunitaire par une proteine virale

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0610046A2 (fr) * 1993-02-01 1994-08-10 Bristol-Myers Squibb Company Vecteurs d'expression codant pour des proteins de fusion bispécifiques et méthodes pour la production de ces protéins dans des cellules de mammifèrres
WO1994021798A1 (fr) * 1993-03-16 1994-09-29 British Technology Group Limited Stimulation d'une reponse immunitaire par une proteine virale

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
C. ERTEL ET AL.: "Viral hemagglutinin augments peptide-specific cytotoxic T cell responses.", EUROPEAN JOURNAL OF IMMUNOLOGY, vol. 23, no. 10, WEINHEIM, DEUTSCHLAND, pages 2592 - 2596 *
H. BOHLEN ET AL.: "Cytolysis of leukemic B-cells by T-cells activated via two bispecific antibodies.", CANCER RESEARCH, vol. 53, no. 18, 15 September 1993 (1993-09-15), BALTIMORE, MD, VSA, pages 4310 - 4314 *
H. BOHLEN ET AL.: "Lysis of malignant B cells from patients with B-chronic lymphocytic leukemia by autologous T cells activated with CD3 x CD19 bispecific antibodies in combination with bivalent CD28 antibodies.", BLOOD, vol. 82, no. 6, 15 September 1993 (1993-09-15), NEW YORK, NY, VSA, pages 1803 - 1812 *
M. FANGER ET AL.: "Bispecific antibodies", CRITICAL REVIEWS IN IMMUNOLOGY, vol. 12, no. 3,4, USA, pages 101 - 124 *
T. NISHIMURA ET AL.: "Human c-erbB-2 proto-oncogene product as a target for bispecific-antibody-directed adoptive tumor immunotherapy.", INTERNATIONAL JOURNAL OF CANCER, vol. 50, no. 5, 12 March 1992 (1992-03-12), GENF, DIE SCHWEIZ, pages 800 - 804 *
V. SCHIRRMACHER: "Active specific immunotherapy - A new modality of cancer treatment involving the patient's own immune system.", ONKOLOGIE, vol. 16, no. 5, GERMERING, DEUTSCHLAND, pages 290 - 296 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1275724A1 (fr) * 2001-07-10 2003-01-15 Volker Prof. Dr. Schirrmacher Réactifs de liaison entre des protéines de surface cellulaire et des cellules effectrices

Also Published As

Publication number Publication date
DE4407538C1 (de) 1995-02-23
JPH09510089A (ja) 1997-10-14
EP0749487A1 (fr) 1996-12-27
US5911987A (en) 1999-06-15

Similar Documents

Publication Publication Date Title
DE60124912T2 (de) Multimerische, einzelkettige, Tandem-Fv-Antikörper
DE3853515T3 (de) Multifunktionelle proteine mit vorbestimmter zielsetzung.
DE69330523T4 (de) Immunoglobuline ohne leichte ketten
DE69922159T2 (de) Mehrzweck-antikörperderivate
DE69633175T2 (de) Multimere proteine
DE69233528T2 (de) Verfahren zur Herstellung von multivalenten antigenbindenden Proteinen
EP1444268B1 (fr) Molecule d'anticorps anti-cd28 bispecifique
DE69133568T2 (de) Bakteriophagenpartikeln,die dAbs präsentieren
DE19742706B4 (de) Lipocalinmuteine
DE19819846B4 (de) Multivalente Antikörper-Konstrukte
EP1206555B1 (fr) Construction d'anticorps fv comportant des sites de liaison pour un recepteur cd16 et une proteine de surface cd30
DE3825615A1 (de) Antigenkonstrukte von "major histocompatibility complex" klasse i antigenen mit spezifischen traegermolekuelen, ihre herstellung und verwendung
DE4014510A1 (de) Variante cd44-oberflaechenproteine, diese kodierende c-dna-sequenzen, antikoerper gegen diese proteine sowie ihre verwendung in der diagnostik und therapie
DE60014575T2 (de) Stabilisierende Peptide, und Polypeptide und Antikörper die diese beinhalten
EP0403961B1 (fr) Conjungués protéiques magnétiques, procédé pour les synthétiser et leur emploi
WO1995024490A1 (fr) Reactif de liaison pour proteine de surface cellulaire et cellule effectrice
EP1307490B1 (fr) Structure fv presentant une affinite influencable avec une substance a lier
EP0537489B1 (fr) Anticorps monoclonaux contre des antigènes associés aux tumeurs, procédé de préparation et utilisation
DE10135039C1 (de) Verfahren zur Isolierung großer Varianzen spezifischer Moleküle für ein Zielmolekül aus Phagemid-Gen-Bibliotheken
EP2600882B1 (fr) ANTICORPS DIRIGÉS CONTRE DES CELLULES DENDRITIQUES HUMAINES 6-SULFO LacNAc-POSITIVES ET UTILISATION DE CEUX-CI
DE60211714T2 (de) Isolierung membrangebundener ligandenspezifischer komplexe
WO1999001475A2 (fr) Anticorps humain diriges contre un (poly)peptide ou une proteine hybride comportant une partie presentant au moins six histidines
EP3201328A1 (fr) Protéine de fusion et procédé de purification
DE4421895A1 (de) Verfahren zur Isolierung von Isolektinen aus der Mistel
EP0115063B1 (fr) Anticorps monoclonaux spécifiques de l'antigène humain du groupe sanguin k (cellano) et lignées cellulaires d'hybridomes produisant ces anticorps monoclonaux

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): JP US

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FR GB GR IE IT LU MC NL PT SE

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
WWE Wipo information: entry into national phase

Ref document number: 1995911308

Country of ref document: EP

WWP Wipo information: published in national office

Ref document number: 1995911308

Country of ref document: EP

WWE Wipo information: entry into national phase

Ref document number: 08702712

Country of ref document: US

WWW Wipo information: withdrawn in national office

Ref document number: 1995911308

Country of ref document: EP